crude cell lysate - storage of crude cell lysate (Apr/14/2006 )

I understand that the best way to store the crude cell lysates for activity assay or western blotting later is to snap freeze it in liquid nitrogen. However, some researchers suggest to store the cell lysates at 4C for a limited period of time. I do not have any first-hand experience about the same. Has somebody tried it before? I would appreciate if somebody shares his/her experience about the later method(i.e. storing at 4C)) with me ?

-maashu-

I personally think it's best to snap and store at -80; 4C is pretty warm and will lead to some problems

are you also adding protease inhibitors to your buffers? and any sort of stabilizer for the activity assay you want to perform? (for example, Na orthovanadate for phosphoproteins, etc)

-aimikins-

QUOTE (aimikins @ Apr 14 2006, 01:34 PM)

I personally think it's best to snap and store at -80; 4C is pretty warm and will lead to some problems

are you also adding protease inhibitors to your buffers? and any sort of stabilizer for the activity assay you want to perform? (for example, Na orthovanadate for phosphoproteins, etc)

Thanks for the response. Yes, I am adding protease inhibitors. I agree with your idea that snap-freezing may be the best.

-maashu-

Hi, i used to freeze proteins by directly keeping them in -80C. if i m looking for activity (MAPK n Rho). i wud snap freeze and store my lysates.may be i can answer ur question in few days. coz i m doing ERK activation experiment today, if i have to repeat the experiment i will do it in few days. as i stored my samples in 4C (since the experiment is just standardisation one).

with best regards

QUOTE (maashu @ Apr 14 2006, 10:22 AM)

QUOTE (aimikins @ Apr 14 2006, 01:34 PM)

I personally think it's best to snap and store at -80; 4C is pretty warm and will lead to some problems

are you also adding protease inhibitors to your buffers? and any sort of stabilizer for the activity assay you want to perform? (for example, Na orthovanadate for phosphoproteins, etc)

Thanks for the response. Yes, I am adding protease inhibitors. I agree with your idea that snap-freezing may be the best.